Preparation And Identification Of Monoclonal Antibody Against Prostate Specific Antigen

Objective To screen paired monoclonal antibody(McAb) with equimolar characteristics against total-prostate specific antigen(t-PSA), with high sensitivity and broad linearity against free-prostate specific antigen(f-PSA). Methods 6-8 weeks BALB/C mice were immunized four times with prostate specific antigen(PSA).Hybridoma cell lines were obtained by fusing mouse myeloma cells and spleen cells of BALB/C mice. Positive clones against PSA were screened by limiting dilution and indirect ELISA using PSA as detecting antigen. Ascites fluid collected by F1 mouse, was then purified, McAbs were then classified by epitopes analysing through indirect ELISA. Paired McAbs were further identified equimolar-response with WHO96/668 and WHO96/668. Results Eighty-eight stable monoclonal hybridoma cell lines were obtained. Six of these which include two pairs of t-PSA McAbs and two pairs of f-PSA McAbs showed high sensitivity, great specifity, broad linearity, and required stablity.Conclusion The paired McAbs screened in this study were good raw material for quantitative analyse t-PSA and f-PSA in human serum, especially suitable for chemiluinescent immunoassay which require high sensitivity and high accuracy.